细粒棘球蚴<em>TSP1</em>和<em>TSP6</em>基因的克隆及生物信息学分析

doi:10.7668/hbnxb.2015.02.008

华北农学报 ›› 2015, Vol. 30 ›› Issue (2): 41-46. doi: 10.7668/hbnxb.2015.02.008

所属专题： 生物技术；

细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

刘田莉¹, 孟庆玲¹, 乔军¹, 陈诚¹, 马玉¹, 胡政香¹, 才学鹏², 陈创夫¹

1. 石河子大学动物科技学院, 新疆石河子 832003;
2. 中国农业科学院兰州兽医研究所, 甘肃兰州 730046

收稿日期:2015-02-10 出版日期:2015-04-28
通讯作者: 孟庆玲(1969-),女,江苏徐州人,教授,博士,主要从事寄生虫分子生物学研究。
作者简介:刘田莉(1992-),女,河南商丘人,在读硕士,主要从事动物寄生虫研究。
基金资助:
国家公益性(农业)行业专项(201303037);国家国际科技合作项目(2014DFR31310);新疆自治区研究生科研创新项目(No.XJGRI2014059)

Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus

LIU Tian-li¹, MENG Qing-ling¹, QIAO Jun¹, CHEN Cheng¹, MA Yu¹, HU Zheng-xiang¹, CAI Xue-peng², CHEN Chuang-fu¹

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2015-02-10 Published:2015-04-28

摘要/Abstract

摘要： 为了研究绵羊细粒棘球蚴重要抗原基因Tetraspanin 1-TSP1(TSP1)和Tetraspanin 1-TSP6(TSP6)的功能,对GenBank中Eg基因组数据库检索,获得 TSP1和TSP6 的cDNA序列并设计特异性引物。以Eg头节为总RNA模板,进行RT-PCR,将PCR产物克隆到pMD19-T载体后测序并进行生物信息学分析。 TSP1 cDNA全长792个核苷酸,编码263个氨基酸,该多肽含有3个潜在的N端糖基化位点,2个N端酰基化位点,与已登录的标准株 TSP1 基因序列(EG_11043)同源性为98.99%,其推导的氨基酸序列同源性为98.48%; TSP6 cDNA全长666个核苷酸,编码221个氨基酸,该多肽含有5个N端酰基化位点,1个酪氨酸激酶磷酸化位点,与已登录的标准株 TSP6 基因序列(EG_00715)同源性为98.18%,其推导的氨基酸序列同源性为85.07%。运用分子生物学软件对Eg TSP1和TSP6 基因进行生物信息学分析,预测已知序列的蛋白质抗原的结构和表位,为疫苗的研发奠定了前期基础。

关键词: 细粒棘球蚴, 抗原基因, 基因克隆, 生物信息学分析

Abstract: In order to study the function of two important antigen genes tetraspanin 1-TSP1(TSP1) and tetraspanin 1-TSP6(TSP6), primers derived from Echinococcus granulosus genome database in GenBank were designed and the open reading frame(ORF) sequences of TSP1 and TSP6 were cloned by RT-PCR from hydatid protoscolex.Then they were cloned into pMD19-T vector for bioinformatics analysis.The results indicated that the TSP1 cDNA contains 792 nucleotides.The deduced protein consisted of 263 amino acids and has three N-glycosylation sites, two N-acylation sites.The gene sequence showed about 98.99% identity with the TSP1 (EG_11043) reported and the induced amino acid sequence showed about 98.48% identity.The TSP6 cDNA contains 666 nucleotides.The deduced protein consisted of 221 amino acids and has five N-acylation sites, one Tyrosine kinase phosphorylation sites.The gene sequence showed about 98.18% identity with the TSP6 (EG_00715) reported and the induced amino acid sequence showed about 85.07% identity.The study carried out bioinformatics analysis of the TSP1 and TSP6 gene of Eg by molecular biology software to predict the structure and epitope of protein antigens known and laid a good foundation for the preparation of developing a vaccine.

Key words: Cystic echinococcosis, Antigen gene, Gene cloning, Bioinformatics analysis

中图分类号:

刘田莉, 孟庆玲, 乔军, 陈诚, 马玉, 胡政香, 才学鹏, 陈创夫. 细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析[J]. 华北农学报, 2015, 30(2): 41-46. doi: 10.7668/hbnxb.2015.02.008.

LIU Tian-li, MENG Qing-ling, QIAO Jun, CHEN Cheng, MA Yu, HU Zheng-xiang, CAI Xue-peng, CHEN Chuang-fu. Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(2): 41-46. doi: 10.7668/hbnxb.2015.02.008.

http://www.hbnxb.net/CN/Y2015/V30/I2/41

参考文献

[1] Li J,Zhang W B,Wilson M,et al.A novel recombinant antigen for immunodiagnosis of human cystic echinococcosis[J].The Journal of Infectious Diseases,2003,188(12):1951-1960.
[2] 陆家海,陈慧红,余新炳,等.细粒棘球蚴全长cDNA文库的构建及初步鉴定[J].中国人兽共患病杂志,2001,17(3):31-33.
[3] 史大中.中国囊性包虫病的地理分布[J].地方病通报,2000,15(1):74-75.
[4] 吴丽文,李凡卡,韩菲,等.2011年新疆兵团人群包虫病流行病学现况调查[J].中国人兽共患病学报,2013,29(4):415-417.
[5] Torgerson P R,Oguljahan B,Muminov A E,et al.Present situation of cystic echinococcosis in Central Asia[J].Parasitology International,2006,55(Suppl):207-212.
[6] 赵莉,张旭,张壮志,等.包虫病诊断技术与预防疫苗的研究进展[J].疾病预防控制通报,2013,28(2):84-87,94.
[7] Chow C,Gauci C G,Cowman A F,et al.Echinococcus granulosus:oncosphere-specific transcription of genes encoding a host-protective antigen[J].Experimental Parasitology,2004,106(3/4):183-186.
[8] Gauci C,Heath D,Chow C,et al.Hydatid disease:vaccinology and development of the EG95 recombinant vaccine[J].Expert Review of Vaccines,2005,4(1):103-112.
[9] Zheng H,Zhang W,Zhang L,et al.The genome of the hydatid tapeworm Echinococcus granulosus[J].Nature Genetics,2013,45(10):1168-1175.
[10] Dang Z,Yagi K,Oku Y,et al.Evaluation of Echinococcus multilocularis tetraspanins as vaccine candidates against primary alveolar echinococcosis[J].Vaccine,2009,27(52):7339-7345.
[11] Marsland B J,Tisdall D J,Heath D D,et al.Construction of a recombinant ORF virus that expresses an Echinococcus granulosus vaccine antigen from a novel genomic insertion site[J].Archives of Virology,2003,148(3):555-562.
[12] Moribe H,Yochem J,Yamada H,et al.Tetraspanin protein (TSP-15) is required for epidermal integrity in Caenorhabditis elegans[J].Journal of Cell Science,2004,117(22):5209-5220.
[13] Cai P,Bu L,Wang J,et al.Molecular characterization of Schistosoma japonicum tegument protein tetraspanin-2:sequence variation and possible implications for immune evasion[J].Biochemical and Biophysical Research Communications,2008,372(1):197-202.
[14] Wright M D,Henkle K J,Mitchell G F.An immunogenic Mr 23 000 integral membrane protein of Schistosoma mansoni worms that closely resembles a human tumor-associated antigen[J].Journal of Immunology (Baltimore,Md.:1950),1990,144(8):3195-3200.
[15] Loukas A,Tran M,Pearson M S.Schistosome membrane proteins as vaccines[J].International Journal for Parasitology,2007,37(3/4):257-263.
[16] Hemler M E.Tetraspanin functions and associated microdomains[J].Nature Reviews Molecular Cell Biology,2005,6(10):801-811.
[17] Berditchevski F.Complexes of tetraspanins with integrins:more than meets the eye[J].Journal of Cell Science,2001,114(23):4143-4151.
[18] Clergeot P H,Gourgues M,Cots J,et al. PLS1,a gene encoding a tetraspanin-like protein,is required for penetration of rice leaf by the fungal pathogen Magnaporthe grisea[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(12):6963-6968.
[19] Levy S,Shoham T.Protein-protein interactions in the tetraspanin web[J].Physiology,2005,20(4):218-224.
[20] Tran M H,Pearson M S,Bethony J M,et al.Tetraspanins on the surface of Schistosoma mansoni are protective antigens against schistosomiasis[J].Nature Medicine,2006,12(7):835-840.

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细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus

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细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus

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